Non-synonymous single nucleotide alterations in the microsomal epoxide hydrolase gene and their functional effects

1. By sequencing genomic DNA from 72 established cell lines derived from Japanese individuals, we detected 25 single nucleotide alterations in the microsomal epoxide hydrolase (EPHX1) gene. Of them, five were exonic alterations resulting in amino acid alterations (77C>G, T26S; 128G>C, R43T; 337T>C, Y113H; 416A>G, H139R; 823A>G, T275A). The T26S, R43T, Y113H and H139R substitutions were found at relatively high frequencies and seemed to be polymorphic, and T26S and T275A were novel. 2. To examine the effects of these amino acid alterations on EPHX1 function, EPHX1 cDNA constructs of wild-type and five variants were transfected into COS-1 cells, and their hydrolytic activities for cis-stilbene oxide were determined in vitro. Although all of the transfectants expressed EPHX1 mRNA and protein at similar levels, the variant H139R protein was expressed at a significantly higher level (128% of the wild-type). K(m) values were not significantly different between the wild-type and variants. 3. Increase (140%) in the enzymatic activity (V(max)) of the variant H139R was accompanied by the increased EPHX1 protein level without any significant change in the intrinsic EPHX1 activity. On the other hand, the variant R43T showed increased values for V(max) and clearance (V(max)/K(m)) (around 130%) both on a microsomal protein basis and on a EPHX1 protein basis. 4. These results suggest that R43T as well as H139R increase epoxide hydrolase activity.     

Antimicrobial activity of hinokitiol against Legionella pneumophila

Antimicrobial activity of hinokitiol (beta-thujaplicin), which is a major component of the essential oil of Chamaecyparis obtuse, against Legionella pneumophila was investigated experimentally. The quantitative antibacterial assay of hinokitiol was carried out by the disk-diffusion method. The test concentrations of hinokitiol were 0.39 to 25.0 micrograms/disk, and the lowest concentrations of hinokitiol that showed growth inhibition against L. pneumophila were 1.56 micrograms/disk on B-SYE agar and 0.39 microgram/disk on B-SYE agar without iron.     

Integration of orosensory and postingestive stimuli for the control of excessive fat intake in mice

OBJECTIVES: Long-term preferences for, and reinforcement effects of, undigested fat substitutes were investigated by means of a two-bottle choice test and the conditioned place preference (CPP) test. METHODS: We tested intact corn oil and sorbitol fatty acid esters, which have been developed as non-digestible fat substitutes with low energy (1.5 kcal/g). RESULTS: Palatability of the sorbitol fatty acid esters was similar to corn oil over 30 min in the short-term two-bottle choice test in mice. However, mice did not continue to eat the fat substitute in the long-term two-bottle choice test, which included postprandial feedback effects. Moreover, sorbitol fatty acid ester did not act as a reinforcer in the CPP test. Mice with 0.1 mL of corn oil placed into their stomachs just before conditioning showed reinforcing effects on taking sorbitol fatty acid ester in the CPP test. However, intragastric administration of corn oil alone without stimulation of the oral cavity did not show any such reinforcing effects against corn oil. CONCLUSIONS: These results suggest that the postingestible effects of corn oil are involved in long-term preference and reinforcing effects.     

Effect of milrinone on vecuronium-induced neuromuscular block

We examined the effect of milrinone, a phosphodiesterase III inhibitor, on neuromuscular block induced by vecuronium. Thirty adult patients were randomly assigned to one of two equal groups: the milrinone group and the control group. Subjects in the milrinone group received an intravenous loading dose of milrinone 5 microg x kg-1x min-1 for 10 min, followed by an infusion at a rate of 0.5 microg x kg-1x min-1. Subjects in the control group received normal saline at a rate of 0.1 ml x kg-1 x h-1. Thirty minutes after the beginning of the infusion of milrinone, anaesthesia was induced with intravenous thiopental 4 mg x kg-1 and fentanyl 2 microg x kg-1, and was maintained with isoflurane in oxygen and nitrous oxide. Neuromuscular blockade was monitored electromyographically at the adductor pollicis muscle. The times from the administration of vecuronium 0.1 mg.kg-1 to the onset of neuromuscular block and the return of the first, second, third, and fourth response of the train-of-four were compared between the two groups. Times to the recovery of the ratio of the first twitch to the control twitch to 25%, 50% and 75%, and times to the recovery of train-of-four ratio to 25%, 50% and 75% were also compared between the two groups. The onset of neuromuscular block in the milrinone group was significantly slower than in the control group. The times to the returns of the four twitches of the train-of-four, times to recovery of the ratio of the first twitch to the control twitch to 25% and 50%, and the times to the recovery of the train-of-four ratio to 25% and 50% were significantly shorter in the milrinone group than in the control group. We conclude that milrinone delays the onset of neuromuscular blockade but hastens its recovery in anaesthetised patients receiving vecuronium.     

Propofol-induced anesthesia in mice is mediated by gamma-aminobutyric acid-A and excitatory amino acid receptors

To elucidate the role of gamma-aminobutyric acid (GABA)(A) receptor complex and excitatory amino acid receptors (N-methyl-D-aspartate [NMDA] and non-NMDA receptors) in propofol-induced anesthesia, we examined behaviorally the effects of GABAergic and glutamatergic drugs on propofol anesthesia in mice. All drugs were administered intraperitoneally. General anesthetic potencies were evaluated using a righting reflex assay. The GABA(A) receptor agonist muscimol potentiated propofol (140 mg/kg; 50% effective dose for loss of righting reflex) induced anesthesia. Similarly, the benzodiazepine receptor agonist diazepam and the NMDA receptor antagonist MK-801 augmented propofol anesthesia, but the non-NMDA receptor antagonist CNQX did not. In contrast, the GABA(A) receptor antagonist bicuculline antagonized propofol (200 mg/kg; 95% effective dose for loss of righting reflex) induced anesthesia. However, neither the benzodiazepine receptor antagonist flumazenil, the GABA synthesis inhibitor L-allylglycine, nor the NMDA receptor agonist NMDA reversed propofol anesthesia. Conversely, the non-NMDA receptor agonist kainate enhanced propofol anesthesia. These results suggest that propofol-induced anesthesia is mediated, at least in part, by both GABA(A) and excitatory amino acid receptors. IMPLICATIONS: We examined behaviorally the effects of GABAergic and glutamatergic drugs on propofol-induced anesthesia in mice. The results suggest that propofol anesthesia is mediated, at least in part, by both GABA(A) and excitatory amino acid receptors.     

Modified Daggett's technique for early repair of postinfarct posterior ventricular septal rupture

A surgical modification for safe early repair of posterior septal rupture is described. This technique is based on the method described by Daggett, but adds one internal patch, plus the application of fibrin glue between the internal and external patch for minimizing bleeding. This modification is a simple and reliable one for repairing posterior ventricular septal rupture.     

Preliminary evaluation of image reconstruction by ordered-subset expectation maximization in thallium-201 lung scintigraphy

OBJECTIVES: We observed whether clearer tumor delineation and greater tumor to non-tumor (T/N) count ratios could be obtained using an iterative ordered-subsets expectation maximization (OSEM) algorithm than conventional filtered-back projection algorithm (FBP) in the image reconstruction of thallium-201 (201Tl) lung scintigraphy. METHODS: In 29 patients with lung cancer and phantom studies, tomograms were reconstructed using FBP and OSEM algorithms, with and without a prefilter (Butterworth filter: BW), whose cut-off frequencies were 0.10 cycles/pixel for FBP and 0.10 and 0.17 cycles/pixel for OSEM. Visual interpretation and tumor to non-tumor (T/N) count ratios were obtained and compared. RESULTS: Without a prefilter, T/N ratios from OSEM and FBP were 1.89 +/- 0.31 (early) and 2.00 +/- 0.54 (late) for OSEM, 1.90 +/- 0.33 (early) and 2.05 +/- 0.59 (late) for FBP, respectively. The OSEM reconstruction without prefiltering showed clearer tumor contours than FBP without a prefilter. Incorporation of BW showed visually low-noised images but decreased T/N ratios in both reconstructions with BW (0.10 cycles/pixel). No greater T/N ratios were obtained by OSEM than FBP, with or without prefiltering. With BW with a cut-off frequency of 0.17 cycles/pixel, the same T/N ratios as those without BW were obtained. The tumor model sized 0.9 cm in the phantom study was invisible in both OSEM and FBP reconstructions without a prefilter, but visible with a prefilter. The influence of prefiltering on T/N ratios was also observed in phantom studies. CONCLUSIONS: Visually improved tumor delineation could be obtained in OSEM reconstruction without a prefilter as compared to FBP reconstruction without a prefilter for tumors greater than 2 cm. Prefiltering should be incorporated into OSEM reconstruction in diagnosing small tumors. However, the influence of prefilter (BW) setting on semi-quantitative interpretation needs further discussion.     

Intractable duodenocutaneous fistula after nephrectomy for stone pyonephrosis: report of a case

A 56-year-old woman was admitted to our hospital for treatment of right stone pyonephrosis with a perirenal abscess. After right nephrectomy for the pyonephrosis, the patient suffered from post-operative bleeding, which was stopped by closing off the drain tube with a clamp. However, a right retroperitoneal abscess with gas formation developed nine days after the operation, necessitating an operative procedure for drainage. Pus culture revealed Staphylococcus epidermidis and Candida albicans. Discharge from the drain tube became dark green days after the drainage procedure. Upper gastrointestinal series revealed a duodenal fistula, which could not be closed using a retroperitoneal approach, so the operative wound was left open. Because of the volume of discharge (800-1,400 ml/day), somatostatin analogue, 100 micrograms, was injected subcutaneously twice a day. Discharge decreased by one-half within 2 weeks of the administration of somatostatin analogue. However, the duodenocutaneous fistula had not resolved over a period of 8 months. Since the patient developed acute cholecystitis, both cholecystectomy and closure of the duodenocutaneous fistula were performed transperitoneally. The duodenocutaneous fistula, which was closed with Endo GIA (35 mm), had protruded from a descending portion of the duodenum like the diverticulum. The postoperative course was uneventful. We speculated that the fistula occurred as a result of the inflammation with the abscess formation.     

The effect of corticosteroids on postoperative course after esophageal resection for carcinoma

BACKGROUND: The effect of steroids against inflammatory mediators is well known, but its benefit and adverse effect on the postoperative clinical course are uncertain after esophageal resection for carcinoma. METHODS: Forty-three cases undergoing esophageal resection for carcinoma were studied retrospectively. Twenty-six cases, given corticosteroids during operation or first postoperative day, were classified as the steroid group, and the other 17 cases were defined as the control group. Postoperative courses, complications, and outcomes were compared between these 2 groups. RESULTS: In the steroid group, heart rate, body temperature and C-response protein were depressed during the postoperative period, and duration of systemic inflammatory response syndrome was 1.2 +/- 1.1 days compared with 2.7 +/- 1.4 days in the control group (P < 0.001). Although the duration of mechanical ventilation and ICU stay was unchanged, morbidity rate of postoperative anastomotic leakage was 15% in the steroid group compared with 47% in the control group (P < 0.05). The other morbidity rates of pulmonary complications, postoperative infection, arrhythmia, and liver dysfunction as well as mortality rate were similar between the 2 groups. CONCLUSIONS: Corticosteroids given in the early postoperative period depress systemic inflammatory response and attenuate the rate of postoperative anastomotic leakage after esophageal resection for carcinoma.     

Resected cases with primary lung cancer after preoperative high dose irradiation

After greater than 60 gray (Gy) irradiation, we performed the pulmonary resection in the 18 primary lung cancer cases. The mean irradiation dose to the tumor was 68.2 (range 60-101) Gy, and the mean irradiation dose to the bronchial stump was 47.1 (range 0-82) Gy. Median time from end of irradiation to surgical resection was 136 (range 20-894) days. One partial resection, 9 single lobectomies, 3 double lobectomies, and 5 pneumonectomies were done. Mainly, we closed the bronchial stump by the automatic stapling device and additional hand suturing. The bronchial stump was covered in the 12 cases by the owner stalk thymus, the intercostals muscular flap, the omentum flap, and et al. The major postoperative complications due to preoperative irradiation were not seen. Bronchopleural fistulas did not occur. Pathologically, the wall of the submucosal capillary vessels were getting thick in the patients operated more than 3 months later after irradiation. In such cases with the decrease of the blood flow, the bronchial stump should be covered. The pulmonary resection after the high dose irradiation was considered to be tolerable.